miR-221-5p regulates proliferation and migration in human prostate cancer cells and reduces tumor growth in vivo
miR-221-5p regulates proliferation and migration in human prostate cancer cells and reduces tumor growth in vivo
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DOI：
10.1186/s12885-019-5819-6
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发表时间：
2019-06-25
期刊：
影响因子：
3.8
通讯作者：
Zoni, Eugenio
中科院分区：
文献类型：
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作者：
Kiener, Mirjam;Chen, Lanpeng;Zoni, Eugenio
尽管前列腺癌(PCA)治疗取得了最新进展，但它仍然是欧洲男性癌症相关死亡的第三大原因。MicroRNAs(MiRNAs)是一种具有基因表达调控功能的非编码小RNA分子，已有报道在所有类型的上皮性肿瘤和血液病中都有报道。方法检索218例PCa患者(GSE21036)的iRNA表达数据，分析miR-221-5p的表达水平。通过miR-221-5p的过表达和敲除实验，研究了miR-221-5p在雄激素依赖和雄激素非依赖的PCa细胞模型(C4-2和PC-3M-Pro4细胞)中的功能作用。通过斑马鱼模型的渗出研究，确定了高侵袭性高表达miR-221-5p的PC-3M-Pro4细胞的转移潜能。最后，通过裸鼠原位移植和肿瘤生长评估，研究miR-221-5p过表达对PC-3M-Pro4Luc2细胞体内生长的影响。结果对人前列腺癌原发和转移标本及对照正常前列腺组织的microRNA表达数据进行分析，发现miR-221-5p在前列腺癌组织中较正常前列腺组织和转移癌组织中显著下调。我们的体外数据表明，miR-221-5p过表达降低了PCa细胞的增殖和集落形成。此外，miR-221-5p的过表达显著减少了PCa细胞的迁移，这与所选EMT标记的差异表达有关。MiR-221-5p过表达的功能变化可通过miR-221-5p水平的降低而逆转，表明miR-221-5p的抑瘤作用是特异性的。此外，在斑马鱼模型中，miR-221-5p的过表达显著减少了PC-3M-Pro4细胞的渗出和转移形成，并在PCa原位小鼠模型中降低了肿瘤负担。结论这些数据有力地支持了miR-221-5p在PCa背景下的肿瘤抑制作用及其作为治疗靶点的潜力。
BackgroundDespite latest advances in prostate cancer (PCa) therapy, PCa remains the third-leading cause of cancer-related death in European men. Dysregulation of microRNAs (miRNAs), small non-coding RNA molecules with gene expression regulatory function, has been reported in all types of epithelial and haematological cancers. In particular, miR-221-5p alterations have been reported in PCa.MethodsmiRNA expression data was retrieved from a comprehensive publicly available dataset of 218 PCa patients (GSE21036) and miR-221-5p expression levels were analysed. The functional role of miR-221-5p was characterised in androgen- dependent and androgen- independent PCa cell line models (C4-2 and PC-3M-Pro4 cells) by miR-221-5p overexpression and knock-down experiments. The metastatic potential of highly aggressive PC-3M-Pro4 cells overexpressing miR-221-5p was determined by studying extravasation in a zebrafish model. Finally, the effect of miR-221-5p overexpression on the growth of PC-3M-Pro4luc2 cells in vivo was studied by orthotopic implantation in male Balb/cByJ nude mice and assessment of tumor growth.ResultsAnalysis of microRNA expression dataset for human primary and metastatic PCa samples and control normal adjacent benign prostate revealed miR-221-5p to be significantly downregulated in PCa compared to normal prostate tissue and in metastasis compared to primary PCa. Our in vitro data suggest that miR-221-5p overexpression reduced PCa cell proliferation and colony formation. Furthermore, miR-221-5p overexpression dramatically reduced migration of PCa cells, which was associated with differential expression of selected EMT markers. The functional changes of miR-221-5p overexpression were reversible by the loss of miR-221-5p levels, indicating that the tumor suppressive effects were specific to miR-221-5p. Additionally, miR-221-5p overexpression significantly reduced PC-3M-Pro4 cell extravasation and metastasis formation in a zebrafish model and decreased tumor burden in an orthotopic mouse model of PCa.ConclusionsTogether these data strongly support a tumor suppressive role of miR-221-5p in the context of PCa and its potential as therapeutic target.